Objectives
Main Objective
The main objective of the project is the development, optimization and implementation of an economically viable process for the integrated management of vinification by-products on a medium pilot scale. The process within the framework of this project refers to the extraction of solid winemaking waste and its subsequent treatment through the development of environmentally friendly techniques, for the isolation and characterization of fractions with the highest anthocyanin content with parallel evaluation of their bioactivity through in vitro studies. In addition, the project aims to the development of all preparatory steps including business and marketing plans, required for the commercialization of novel formulations enriched in anthocyanins.
Specific Objectives-Methodology
Winery by-products collection and processing
Pomace resulting from the vinification of different grape varieties, both domestic and imported (Mavrotragano, Syrah, Merlot), represents the raw material for anthocyanins extraction and is supplied within the context of established collaborations with wineries. Our objective is to process pomace through the development of low cost extraction protocols, suitable for anthocyanins extraction, that can be applied on a larger scale. Utilizing materials generally recognized as safe (GRAS), we ensure that the acquired anthocyanin-rich extracts are suitable and safe for the subsequent development of food additives, functional foods and /or pharmaceuticals.
The methodology utilized to achieve this goal entails pomace drying and pulverizing, and subsequent extraction based on the use of ultrasounds and ethyl alcohol, followed by condensation and solvent recycling for reuse.
Optimization and standardization of analytical methods for the quality control of extracts produced through large scale applications
Our objective is to apply the extraction method in medium/large scale and to concomitantly develop analytical protocols for the identification of bioactive compounds in the acquired extracts, volatile and non-volatile, aiming to the qualitative and quantitative characterization of the extracts and the determination of fractions presenting the highest anthocyanin concentration. This will allow obtaining quantified extracts suitable for further, processing, formulation and use.
The methodology used refers to the optimization and standardization of the extraction process for use in medium/large scale entailing the processing of 5 kg raw material and the development and verification of analytical methods (HPLC-DAD-MS, GC-MS and NMR), for the identification of the bioactive compounds included in the acquired extracts.
Identification and characterization of extracts enriched in bioactive compounds (anthocyanin) exerting strong antioxidant effects in in vitro systems representative of neurodegenerative disorders
Our objective is to further characterize selected, anthocyanin-rich extracts, in terms of their bioactivity, utilizing in vitro systems representative of neurodegenerative disorders and molecular methods for assessing their antioxidant effects, their ability to stimulate molecular pathways that enhance cellular survival and their ability to reduce/inhibit or alleviate protein aggregation.
The methodology used for achieving this goal refers to molecular assays conducted in neuronal cell lines (murine neuroblastoma, N2a, and human neuroblastoma, SH-SY5Y), and includes: a) cytotoxicity testing of the selected extracts, entailing determination of IC50 values and determination of cell viability (flow cytometry) following incubation with different concentrations of the extracts under study, b) assessment of the cellular metabolism and mitochondrial function (antioxidant potential, glycolytic flux), to determine whether the tested extracts enhance the cellular antioxidant potential, resulting in reduced reactive oxygen species (ROS) levels and c) assessment of their ability to stimulate signalling pathways that promote cell survival . To evaluate the anti-aggregation effects of the extracts under study, we utilize one of the most widely used cellular models of prion diseases, corresponding to murine neuroblastoma cells persistently infected with prion (scN2a). This cellular model displays accumulation and aggregation of the disease-associated PrPSc protein, which corresponds to the aberrantly folded isoform of the normal PrPC protein. Utilizing this model we determine whether the tested extracts induce dose-related reduction of PrPSc levels.
Feasibility study and development of commercial activities for the introduction, promotion and commercialization of novel, anthocyanin-rich formulations with verified neuroprotective and antineoplastic effects
Our objective is to conduct a thorough study which will include a market study, the development of a financial plan and of a marketing plan, aiming to commercialize novel, anthocyanin-rich products.
Our approach towards this direction entails: a) a thorough study of the local and European market and the evaluation of benefits, advantages, disadvantages, opportunities and risks (SWOT), with the aim to achieve the best possible strategic planning of commercialization, b) the estimation of the costs referring to each individual step of the production process and of the final product, c) the estimation of anticipated sales and the development of a financial plan, d) the development of a marketing plan, focusing on the product, the consumers, the selection of the best distribution sites and the product price.
Model of development and commercialization of novel, anthocyanin-rich products
Our objective is to complete all the required contacts with regulatory agencies (EOF, EFSA) and proceed to the submission of a health claim application to EFSA.